Last data update: 27 January 2021 04:25 CET
Plasmid name: pCAGGS-FLAG-TAX1BP1-L (LMBP 5163)
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|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
Human T-cell leukemia virus type I (HTLV-1) Tax1 binding protein 1 cDNA (TXBP15-1; TXBP151; TAX1BP1); large form (TAX1BP1-L)|
FLAG epitope tag; N-terminal
|Promoter:||Chicken β-actin/rabbit β-globin hybrid promoter (AG)
Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only
Escherichia coli lac operon promoter
|Terminator:||Rabbit β-globin polyadenylation signal (β-globin polyA)
Simian virus 40 polyadenylation signal (SV40 polyA)
|Selection marker:||Ampicillin (amp)|
|Replicon:||Escherichia coli plasmid pMB1 origin
Simian virus 40 bidirectional origin (SV40)
|Host range:||Escherichia coli
Mammalian cells; SV40 permissive cells
|Parental clone:||pCAGGS-FLAGmA20; pCAGGS-TAX1BP1-L|
|Further information:||The plasmid was constructed as follows: 1) The N-terminal domain of the human TAX1BP1-L was generated by PCR. 2) The C-terminal domain was cut out of pCAGGS-TAX1BP1-L with ScaI and XhoI. 3) Both fragments were ligated behind the FLAG epitope into the KpnI-SalI opened pCAGGS-FLAGmA20 plasmid.
The plasmid is useful for highly efficient expression of human TAX1BP1-L under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells.
When cloning a fragment downstream from the lac promoter it may be advisable to use lacIq strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727098.1.
According to the depositor, the human TAX1BP1-L CDS is a slightly shorter splice variant compared to the human TAX1BP1-L2 CDS of the plasmid pCAGGS-FLAG-TAX1BP1-L2 (LMBP 5665). According to sequencing results however, the only difference with pCAGGS-FLAG-TAX1BP1-L2 is that pCAGGS-FLAG-TAX1BP1-L contains an additional fragment of pCAGGS upstream of the 3' UTR of the rabbit β-globin gene.
The entire human TAX1BP1 CDS was sequenced at BCCM/GeneCorner.
The nucleotide sequence of the human TAX1BP1 coding sequence corresponds with the Genbank accession number NM_001206901.1, except for an A instead of T at nucleotide position 919 of the human TAX1BP1 CDS, resulting in a isoleucine instead of leucine at amino acid position 307. As compared to the EMBL accession number U33821.2, there are more nucleotide differences.
Other name of the plasmid is pCAGGS-flag-TXBP151.
|EMBL Accession number:||NM_001206901.1, view at GenBank
LT727098.1, view at EMBL, GenBank, DDBJ
|Latest sequence update:||16/07/2014|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BglII/XhoI, EcoRI, SpeI and XbaI/XmnI.|
|History of deposit:||This plasmid was deposited by L. Verstrepen(1) (2) and Prof. Dr R. Beyaert(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 MC1061|
|Host reference:||Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
|Related host reference:||Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml)|
|Cultivation remark:||Take a very low plasmid yield into account while choosing a plasmid DNA isolation procedure.|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pCAGGS-FLAG-TAX1BP1-L (LMBP 5163) is available at BCCM/GeneCorner. This plasmid was deposited by L. Verstrepen and Prof. Dr R. Beyaert .|
Note: Up-to-date, validated data are enclosed with the biological material. Nevertheless, these data are a snapshot at a given moment; further updates are always possible.